RESUMEN
Rapid and accurate diagnostic tests are fundamental for improving patient outcomes and combating infectious diseases. The Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) Cas12a-based detection system has emerged as a promising solution for on-site nucleic acid testing. Nonetheless, the effective design of CRISPR RNA (crRNA) for Cas12a-based detection remains challenging and time-consuming. In this study, we propose an enhanced crRNA design system with deep learning for Cas12a-mediated diagnostics, referred to as EasyDesign. This system employs an optimized convolutional neural network (CNN) prediction model, trained on a comprehensive data set comprising 11,496 experimentally validated Cas12a-based detection cases, encompassing a wide spectrum of prevalent pathogens, achieving Spearman's ρ = 0.812. We further assessed the model performance in crRNA design for four pathogens not included in the training data: Monkeypox Virus, Enterovirus 71, Coxsackievirus A16, and Listeria monocytogenes. The results demonstrated superior prediction performance compared to the traditional experiment screening. Furthermore, we have developed an interactive web server (https://crispr.zhejianglab.com/) that integrates EasyDesign with recombinase polymerase amplification (RPA) primer design, enhancing user accessibility. Through this web-based platform, we successfully designed optimal Cas12a crRNAs for six human papillomavirus (HPV) subtypes. Remarkably, all the top five predicted crRNAs for each HPV subtype exhibited robust fluorescent signals in CRISPR assays, thereby suggesting that the platform could effectively facilitate clinical sample testing. In conclusion, EasyDesign offers a rapid and reliable solution for crRNA design in Cas12a-based detection, which could serve as a valuable tool for clinical diagnostics and research applications.
RESUMEN
Organic-inorganic hybrid perovskite quantum dots (QDs) have garnered significant research interest owing to their unique structure and optoelectronic properties. However, their poor optical performance in ambient air remains a significant limitation, hindering their advancement and practical applications. Herein, three amino acids (valine, threonine and cysteine) were chosen as surface ligands to successfully prepare highly luminescent CH3NH3PbBr3 (MAPbBr3) QDs. The morphology and XRD results suggest that the inclusion of the amino acid ligands enhances the octahedral structure of the QD solutions. Moreover, the observed blue-shifted phenomenon in the photoluminescence (PL) aligns closely with the blue-shifted phenomenon observed in the ultraviolet-visible (UV-Vis) absorption spectra, attributed to the quantum confinement effect. The time-resolved spectra indicated that the introduction of the amino acid ligands successfully suppressed non-radiative recombination, consequently extending the fluorescence lifetime of the MAPbBr3 QDs. The photoluminescence quantum yields (PLQYs) of the amino acid-treated MAPbBr3 QDs are increased by 94.8%. The color rendering index (CRI) of the produced white light-emitting diode (WLED) is 85.3, with a correlated color temperature (CCT) of 5453 K. Our study presents a novel approach to enhancing the performance of perovskite QDs by employing specially designed surface ligands for surface passivation.
RESUMEN
BACKGROUND: The pregnant women with intrahepatic cholestasis were at high risk of fetal distress, preterm birth and unexpected stillbirth. Intrahepatic cholestasis of pregnancy (ICP) was mainly caused by disorder of bile acid metabolism, whereas the specific mechanism was obscure. METHODS: We performed proteomics analysis of 10 ICP specimens and 10 placenta specimens from patients without ICP through data-independent acquisition (DIA) technique to disclose differentially expressed proteins. We executed metabolomic analysis of 30 ICP specimens and 30 placenta specimens from patients without ICP through UPLC-MS/MS to identify differentially expressed metabolites. Enrichment and correlation analysis was used to obtain the direct molecular insights of ICP development. The ICP rat models were constructed to validate pathological features. RESULTS: The heatmap of proteomics analysis showed the top 30 up-regulated and 30 down-regulated proteins. The metabolomic analysis revealed 20 richer and 4 less abundant metabolites in ICP samples compared with placenta specimens from patients without ICP, and enrichment pathways by these metabolites included primary bile acid biosynthesis, cholesterol metabolism, bile secretion, nicotinate and nicotinamide metabolism, purine metabolism and metabolic pathways. Combined analysis of multiple omics results demonstrated that bile acids such as Glycohyocholic acid, Glycine deoxycholic acid, beta-Muricholic acid, Noncholic acid, cholic acid, Gamma-Mercholic Acid, alpha-Muricholic acid and Glycochenodeoxycholic Aicd were significantly associated with the expression of GLRX3, MYL1, MYH7, PGGT1B, ACTG1, SP3, LACTB2, C2CD5, APBB2, IPO9, MYH2, PPP3CC, PIN1, BLOC1S1, DNAJC7, RASAL2 and ATCN3 etc. The core protein ACAT2 was involved in lipid metabolic process and animal model showed that ACAT2 was up-regulated in placenta and liver of pregnant rats and fetal rats. The neonates had low birth weight and Safranin O-Fast green FCF staining of animal models showed that poor osteogenic and chondrogenic differentiation of fetal rats. CONCLUSION: Multiple metabolites-alpha-Muricholic acid, beta-Muricholic acid, Glycine deoxycholic acid and Glycochenodeoxycholic Acid etc. were perfect biomarkers to predict occurrence of ICP. Bile acids were significantly associated with varieties of protein expression and these proteins were differentially expressed in ICP samples. Our study provided several biomarkers for ICP detection and potential therapeutic targets for ICP development.
Asunto(s)
Ácidos y Sales Biliares , Biomarcadores , Colestasis Intrahepática , Metabolómica , Placenta , Complicaciones del Embarazo , Proteómica , Femenino , Colestasis Intrahepática/metabolismo , Colestasis Intrahepática/diagnóstico , Humanos , Embarazo , Complicaciones del Embarazo/metabolismo , Complicaciones del Embarazo/diagnóstico , Biomarcadores/metabolismo , Biomarcadores/análisis , Proteómica/métodos , Ácidos y Sales Biliares/metabolismo , Ratas , Placenta/metabolismo , Animales , Metabolómica/métodos , Adulto , Modelos Animales de Enfermedad , Espectrometría de Masas en TándemRESUMEN
Subsequently to the publication of the above article, an interested reader drew to the authors' attention that certain of the EdU assay data shown in Fig. 7E on p. 2418 had already appeared in different form in a previously published paper written by different authors at different research institutes. Owing to the fact that the contentious data in the above article had already been published prior to its submission to International Journal of Oncology, the Editor has decided that this paper should be retracted from the Journal. The authors were asked for an explanation to account for these concerns, but the Editorial Office did not receive a reply. The Editor apologizes to the readership for any inconvenience caused. [International Journal of Oncology 53: 24092422, 2018; DOI: 10.3892/ijo.2018.4586].
RESUMEN
BACKGROUND: Prenatal inflammation exposure (PIE) can increase the disease susceptibility in offspring such as lung cancer. Our purpose was to investigate the mechanisms of PIE on lung cancer. METHODS: Prenatal BALB/c mice were exposed to lipopolysaccharide (LPS), and then, their offspring were intraperitoneally instilled with urethane to establish the two-stage lung cancer carcinogenesis model. At the 48 weeks of age, the offspring mice were killed and lung tissues were collected for HE, immunohistochemistry, immunofluorescence, and Luminex MAGPIX®-based assays. CD11b + F4/80 + tumor-associated macrophages (TAMs) were sorted out from lung tumor tissues by cell sorting technique. Flow cytometry was employed to evaluate the extent of M2-like polarization of TAMs and PD-L1 expression. RESULTS: The offspring of PIE mice revealed more lung lesion changes, including atypical hyperplasia and intrapulmonary metastases. The number of lung nodules, lung organ index, and PCNA, MMP-9 and Vimentin positive cells in lung tissue of PIE group were higher than those of Control group. The increases of mRNA encoding M2 macrophage markers and cytokines in offspring of prenatal LPS-treated mice confirmed the induced effect of PIE on macrophage polarization. Additionally, PIE treatment increased the percentage of CD163 + CD206 + cells in the sorted TAMs. Importantly, endoplasmic reticulum (ER) stress-markers like GRP78/BIP and CHOP, p-IRE1α and XBP1s, and PD-L1 were up-regulated in TAMs from PIE group. Besides, we also observed that IRE1α inhibitor (KIRA6) reversed the M2-like TAMs polarization and metastasis induced by PIE. CONCLUSIONS: IRE1α/XBP1-mediated M2-like TAMs polarization releases the pro-tumorigenic cytokines and PD-L1 expression, which may be the regulatory mechanism of accelerating lung cancer in offspring of mice undergoing PIE.
Asunto(s)
Neoplasias Pulmonares , Animales , Ratones , Neoplasias Pulmonares/patología , Endorribonucleasas/metabolismo , Endorribonucleasas/farmacología , Macrófagos Asociados a Tumores/metabolismo , Antígeno B7-H1/metabolismo , Lipopolisacáridos/farmacología , Transducción de Señal , Proteínas Serina-Treonina Quinasas/metabolismo , Carcinogénesis , Citocinas , Inflamación , Microambiente Tumoral/genéticaRESUMEN
Many metastatic cancers with poor prognoses correlate to downregulated CD82, but exceptions exist. Understanding the context of this correlation is essential to CD82 as a prognostic biomarker and therapeutic target. Oral squamous cell carcinoma (OSCC) constitutes over 90% of oral cancer. We aimed to uncover the function and mechanism of CD82 in OSCC. We investigated CD82 in human OSCC cell lines, tissues, and healthy controls using the CRISPR-Cas9 gene knockout, transcriptomics, proteomics, etc. CD82 expression is elevated in CAL 27 cells. Knockout CD82 altered over 300 genes and proteins and inhibited cell migration. Furthermore, CD82 expression correlates with S100 proteins in CAL 27, CD82KO, SCC-25, and S-G cells and some OSCC tissues. The 37-50 kDa CD82 protein in CAL 27 cells is upregulated, glycosylated, and truncated. CD82 correlates with S100 proteins and may regulate their expression and cell migration. The truncated CD82 explains the invasive metastasis and poor outcome of the CAL 27 donor. OSCC with upregulated truncated CD82 and S100A7 may represent a distinct subtype with a poor prognosis. Differing alternatives from wild-type CD82 may elucidate the contradictory functions and pave the way for CD82 as a prognostic biomarker and therapeutic target.
Asunto(s)
Carcinoma de Células Escamosas , Neoplasias de la Boca , Humanos , Neoplasias de la Boca/patología , Carcinoma de Células Escamosas/metabolismo , Proteína Kangai-1/metabolismo , Tetraspaninas/metabolismo , Proteínas S100 , Biomarcadores , Proteína A7 de Unión a Calcio de la Familia S100RESUMEN
Background: Wuhu Oral Liquid (WHOL) is a modified preparation derived from the famous Wuhu Powder, which has a long history of use in treating traumatic injuries. This preparation has anti-inflammatory and analgesic properties and accelerates recovery following acute soft tissue injuries. Aims: To evaluate the efficacy and safety of WHOL in treating acute soft tissue injury associated with qi stagnation and blood stasis syndrome and to provide a basis for applying for the protection of varieties of Chinese medicine for WHOL. Methods: This study was a randomized, controlled, double-blind, multicenter clinical trial in which Fufang Shang Tong Capsule (FFSTC) was selected as the control drug. A total of 480 subjects with acute soft tissue injury associated with qi stagnation and blood stasis syndrome were randomly divided into a test and control group in a 3:1 ratio. The duration of drug treatment was 10 days. The primary outcome was Visual Analogue Scale (VAS) score for pain (including pain at rest and pain on activity). Secondary outcomes included the disappearance time of the pain at rest and on activity; the curative effect of TCM syndrome and improvement in the individual symptoms of TCM (swelling, ecchymosis, and dysfunction); and changes in C-reactive protein (CRP) and interleukin-6 (IL-6) levels. Safety was assessed using vital signs, laboratory examinations, electrocardiograms, and physical examinations. Results: Patient compliance was satisfactory in both groups (all between 80% and 120%). After 4 days of treatment, the WHOL group was superior to the FFSTC group in decreasing the VAS scores for pain at rest (-1.88 ± 1.13 vs. -1.60 ± 0.93, p < 0.05) and on activity (-2.16 ± 1.18 vs. -1.80 ± 1.07, p < 0.05). After 7 days of treatment, the WHOL group was superior to the FFSTC group in decreasing the VAS scores for pain on activity (-3.87 ± 1.60 vs. -3.35 ± 1.30, p < 0.01) and improving swelling (cure rate: 60.4% vs. 46.2%, p < 0.05; obvious effective rate: 60.7% vs. 47.0%, p < 0.05). After 10 days of treatment, the WHOL group was superior to the FFSTC group in decreasing the levels of CRP (-0.13 ± 2.85 vs. 0.25 ± 2.09, p < 0.05) and improving the TCM syndrome (cure rate: 44.1% vs. 30.8%, p < 0.05) and swelling (cure rate: 75.6% vs. 67.5%, p < 0.01; obvious effective rate: 75.6% vs. 68.4%, p < 0.05; effective rate: 77.0% vs. 71.8%, p < 0.05). The disappearance time of pain at rest was 8 days in both groups and 9 days on activity in both groups. In addition, there was no statistical difference between the incidence of adverse events (4.5% vs. 2.6%, p > 0.05) and adverse reactions (0.3% vs. 0%, p > 0.05) between the WHOL group and the FFSTC group. No serious adverse events occurred in either group, and no subjects were withdrawn because of adverse events. Conclusion: WHOL relieves the symptoms caused by acute soft tissue injury associated with qi stagnation and blood stasis syndrome more rapidly than FFSTC, and it is effective and safe in the treatment of acute soft tissue injury. Future studies still need a larger sample size to verify its efficacy and safety. Clinical Trial Registration: https:// www.chictr.org.cn/showproj.html?proj=149531, Identifier ChiCTR2200056411.
RESUMEN
Clastic rock aquifer of the coal seam roof often constitutes the direct water-filling aquifer of the coal seam and its water-richness is closely related to the risk of roof water inrush. Therefore, the evaluation of the water-richness of clastic rock aquifer is the basic work of coal seam roof water disaster prevention. This article took the 4th coal seam in Huafeng mine field as an example. It combined the empirical formula method and generalized regression neural network (GRNN) to calculate the development height of water-conducting fracture zone, determined the vertical spatial range of water-richness evaluation. Depth of the sandstone floor, brittle rock ratio, lithological structure index, fault strength index, and fault intersections and endpoints density were selected as the main controlling factors. A combination weighting method based on the analytic hierarchy process (AHP), rough set theory (RS), and minimum deviation method (MD) was proposed to determine the weight of the main controlling factors. Introduced the theory of unascertained measures and confidence recognition criteria to construct an evaluation model for the water-richness of clastic rock aquifers, the study area was divided into three zones: relatively weak water-richness zones, medium water-richness zones, and relatively strong water-richness zones. By comparing with the water inrush points and the water inflow of workfaces, the evaluation model's water yield zoning was consistent with the actual situation, and the prediction effect was good. This provided a new idea for the evaluation of the water-richness of the clastic rock aquifer on the roof of the mining coal seam.
RESUMEN
When Chinese coal mines are mining Carboniferous Permian coal seams, the mechanism of water inrush from the roof of the working face usually conforms to the "Upper Three Zones" or "Upper Four Zones" theory. The water inrush passageway is water-conducting fracture zone, and the water inrush position is located in the goaf. However, when mining Jurassic coal seams in Chinese coal mines, the location of water inrush often appears at the head-on working face, above the coal mining machine. Due to the support of the fully mechanized mining support, the roof rock layer cannot collapse and therefore cannot form water-conducting fracture zone. Therefore, the water inrush mechanism cannot be explained by the above two theories. This paper is guided by the Practical Mine Pressure Control Theory, and based on the explanation of the motion forms of bending (pulling) failure movement and shearing (cutting) failure movement, and combined with on-site examples, it is revealed that the passageways leading to from the head-on working face roof in a Jurassic coal seam in the Ordos Basin are splitting zones type, fracture line type and structural fracture type, respectively. Taking the changes in water inflow during the mining process of the 3301 and 3302 working faces in Zhujiamao Coal Mine as examples, this paper reveals the mechanism of water inrush from the head-on working face roof caused by splitting zones type, and proves the existence of this passageway through on-site 3D high-density electrical detection and tracing experiments. Taking two catastrophic water inrush accidents that occurred head-on in the 1309 working face of Guojiahe Coal Industry Co., Ltd. as examples, the water inrush mechanism of the fracture line type and the water inrush mechanism of the structural fracture type were respectively revealed. Based on mechanism of water inrush from head-on roof of working face and the analysis of the on-site water inrush process, a method for distinguishing the type of water inrush passageway from the front roof of the working face is proposed. The results indicate that the Jurassic coal seam mining in the Ordos Basin is prone to shearing (cutting) failure movement, resulting in the frequent formation of the three types of water inrush passageways mentioned above.
RESUMEN
Objective:To evaluate the outcomes following the use of folded supraclavicular flap repaired hypopharyngeal combined neck skin defects. Methods:Folded supraclavicular flap were designed to repaired four cases of hypopharyngeal combined neck skin defects patients. Evaluate their repair effect ï¼3 men,1 woman; mean age:66 yearsï¼. Hypopharyngeal fistula from 3.2 cm×1.8 cm to 4.5 cm×3.8 cm,skin defects in the neck from 3.8 cm×2.2 cm to 5.1 cm×4.5 cm. The folded supraclavicular flap from 8 cm×5 cm to 13 cm×9 cm. Results:All flaps survived, head and neck appearance was satisfactory, and no patient experienced a major complication. All patients resumed an oral diet function. No fistula recurrence or stricture developed after 6-19 months of follow-up. Conclusion:The folded supraclavicular flap is a safe and effective flap Repaired for hypopharyngeal combined neck skin defects.
Asunto(s)
Fístula , Procedimientos de Cirugía Plástica , Traumatismos de los Tejidos Blandos , Masculino , Femenino , Humanos , Anciano , Colgajos Quirúrgicos , Cuello/cirugía , Trasplante de Piel , Cabeza/cirugía , Fístula/cirugía , Traumatismos de los Tejidos Blandos/cirugía , Resultado del TratamientoRESUMEN
The use of gaseous CO in Pd-catalyzed carbonylative quinolone synthesis presents challenges related to safety and precise pressure control. In response, a streamlined non-gaseous synthesis of 4-quinolone compounds has been developed. This study introduces a tunable CO-releasing system utilizing Fe(CO)5 activated by a dual-base system of piperazine and triethylamine. This alternative liquid CO resource facilitates the palladium-catalyzed carbonylative C-C coupling and subsequent intramolecular cyclization. By tuning the tandem kinetics of carbonylation and cyclization, this non-gaseous method achieves the successful synthesis of 22 distinct 4-quinolones with excellent yields. This is achieved through the three-component condensation of sub-stoichiometric amounts of Fe(CO)5 with 2-iodoaniline and terminal alkynes. Operando mechanistic studies have revealed a novel CO transfer mechanism that facilitates homogeneous carbonylative cyclization, distinguishing this method from traditional techniques. In addition to addressing safety concerns, this approach also provides precise control over selectivity, with significant implications for pharmaceutical research and the efficient synthesis of pharmaceutical and bioactive compounds.
RESUMEN
China is an extremely water-scarce country with an uneven distribution of regional water resources. We define two absolute sustainability indicators, using the multi-regional inputâoutput (MRIO) model to outline the contribution of China's physical and virtual water transfers in mitigating the problem of regional water boundary-exceeding. Although the overall use of freshwater resources is within the safe operation space, 55% of province's water resource development transgresses the local water planetary boundary. Physical and virtual water transfers effectively mitigate the stress of water supply to the water planetary boundary in China's water-scarce regions. Among them, the role of virtual water transfers occupies the main part. The cost of using physical water in water-receiving regions and the situation of virtual water flowing from water-scarce regions to developed water-rich regions cannot be ignored, and a small number of provinces are responsible for most of the virtual water net imports and exports. The obtained results are helpful for the redistribution of water planetary boundary transgressing responsibilities among provinces and the formulation of absolute sustainable water resources management policies.
Asunto(s)
Abastecimiento de Agua , Agua , Recursos Hídricos , China , Agua DulceRESUMEN
Achieving both high sensitivity and wide detecting range is significant for the applications of triboelectric nanogenerator-based self-powered pressure sensors (TPSs). However, most of the previous designs with high sensitivity usually struggle in a narrow pressure detection range (<30 kPa) while expanding the detection range normally sacrifices the sensitivity. To overcome this well-known obstacle, herein, piezopotential enhanced triboelectric effect realized by a rationally designed PDMS/ZnO NWs hierarchical wrinkle structure was exploited to develop a TPS (PETPS) with both high sensitivity and wide detecting range. In this PETPS design, the piezopotential derived from the deformation of ZnO NWs enhances its tribo-charge transferring ability; meanwhile, the hierarchical structure helps to establish a dynamically self-adjustable contact area. Benefiting from these advantages, the PETPS simultaneously achieves high sensitivity (0.26 nC cm-2 kPa-1 from 1 to 25 kPa, and 0.02 nC cm-2 kPa-1 from 25 to 476 kPa), fast response (46 ms), wide sensing range (1 to 476 kPa), and good stability (over 4000 cycles). In addition, the output charge density that is independent of the speed rate of driven force was adopted as the sensing signal of PETPS to replace the commonly used peak voltage/current values, enabling it more adaptive to accurately detect pressure variation in real applications.
RESUMEN
Electrocardiogram (ECG) is a widely used technique for diagnosing cardiovascular disease. The widespread emergence of smart ECG devices has sparked the demand for intelligent single-lead ECG-based diagnostic systems. However, it is challenging to develop a single-lead-based ECG interpretation model for multiple disease diagnosis due to the lack of some key disease information. We aim to improve the diagnostic capabilities of single-lead ECG for multi-label disease classification in a new teacher-student manner, where the teacher trained by multi-lead ECG educates a student who observes only single-lead ECG We present a new disease-aware Contrastive Lead-information Transferring (CLT) to improve the mutual disease information between the single-lead-based ECG interpretation model and multi-lead-based ECG interpretation model. Moreover, We modify the traditional Knowledge Distillation into Multi-label disease Knowledge Distillation (MKD) to make it applicable for multi-label disease diagnosis. The whole knowledge transferring process is inter-lead Multi-View Knowledge Transferring of ECG (MVKT-ECG). By employing the training strategy, we can effectively transfer comprehensive disease knowledge from various views of ECG, such as the 12-lead ECG, to a single-lead-based ECG interpretation model. This enables the model to extract intricate details from single-lead ECG signals and enhances the model's capability of diagnosing and identifying single-lead signals. Extensive experiments on two commonly used public multi-label datasets, ICBEB2018 and PTB-XL demonstrate that our MVKT-ECG yields exceptional diagnostic performance improvements for single-lead ECG. The student outperforms its baseline observably on the PTB-XL dataset (1.3 % on PTB.super, and 1.4 % on PTB.sub), and on ICBEB2018 dataset (3.2 %).
RESUMEN
Atmospheric precipitation floods mining areas, which seriously affects the safe production of coal mines. However, research on the mechanism underlying precipitation supplying coal mines, particularly in karst landform areas, remains far from sufficient. Based on the collection of a large amount of geological and hydrogeological mining data and some data related to mine atmospheric precipitation and mine water inflow, the channels of atmospheric precipitation supplying mines in the main coal-producing areas of Guizhou, China, are systematically analysed and studied. They are divided into weathering zone fractures, mining fractures, water diversion faults, water diversion collapse columns and karst channels. Recharge channels have the characteristics of surface infiltration, pipeline flow and layered recharge, as well as self-healing after being filled by surface loess and other materials. The supply of atmospheric precipitation to the coal mine stope is seasonal. The mine water inflow in the rainy season is 1.2 ~ 12 times that in the dry season, with an average of 1.9 times. The supply has hysteresis. The lag time of surface infiltration, pipeline flow and layered flow is 2 ~ 4 days, within 24 h and more than 2 days, respectively. The recharge is affected by the burial depth of the coal seam and the characteristics of the combined upper roof slate. Among the mines affected by atmospheric precipitation and water disasters, some mines have carried out research on the comprehensive treatment of water disasters, implemented supplementary exploration projects such as surface hydrogeological drilling and geophysical exploration, or carried out hydrochemical research. Some mines have adopted water prevention and control projects, such as blocking ground water diversion cracks, constructing water diversion projects, adjusting the mining time of the working face, transforming the drainage system and improving the drainage capacity, to ensure the safe production of mines. This research achievement may provide a theoretical basis and practical experience for the prevention and control of atmospheric precipitation infiltration in coal mines in karst areas.
RESUMEN
Quercus mongolica is a common landscape, afforestation, and construction timber species in northern China with high ecological, economic, and ornamental value. Leaf senescence is a complex process that has important implications for plant growth and development. To explore changes of metabolites during the ageing of Quercus mongolica leaves, we investigated physiological responses and metabolite composition in ageing leaves harvested from 15-20-year-old Quercus mongolica. Leaf samples of Q. mongolica were collected when they were still green (at maturity) (stage 1), during early senescence (stage 2), and during late senescence (stage 3). These leaves were then subjected to physiological index and metabolome sequencing analyses. The physiological analysis showed that the leaves of Q. mongolica changed from green to yellow during senescence, which induced significant accumulation of soluble sugar and significant reductions in the concentration of soluble protein and chlorophyll. Peroxidase and catalase were the main antioxidant enzymes mitigating leaf senescence. Metabolomic analysis identified 797 metabolites during leaf senescence. Compared to stage 1, 70 differential metabolites were screened in stage 2 and 72 were screened in stage 3. Differential metabolites in the two senescent stages were principally enriched in amino acid metabolism, lipid metabolism and secondary metabolite biosynthesis. The contents of N-oleoylethanolamine and N, N-dimethylglycine were significantly increased only in stage 2, while the contents of trifolin, astragalin, valine, isoleucine, leucine, and citric acid were significantly increased only in stage 3. Histidine, homoserine, tryptophan, tyrosine, phenylalanine, proline, norleucine, N-glycyl-L-leucine, linoleic acid, linolenic acid, gallic acid, 3-indoleacrylic acid, 3-amino-2-naphthoic acid, 3-hydroxy-3-methylpentane-1,5-dioic acid, 2,3,4-trihydroxybenzoic acid, trifolin, astragalin, DL-2-aminoadipic acid, pinoresinol dimethyl ether, dimethylmatairesinol, and lysophosphatidylcholine increased during both stage 2 and stage 3. Increasing contents of these metabolites may constitute the main mechanism by which Q. mongolica leaves adapt to senescence.
Asunto(s)
Quercus , Metabolómica , Aminoácidos , Metaboloma , PeroxidasasRESUMEN
Background: Non-small cell lung cancer (NSCLC) is a common lung tumor with high mortality. The complex formed by MYB-MuvB complex (MMB) and forkhead box M1 (FOXM1) (MMB-FOXM1) plays a vital role in cell cycle progression to affect the progression of diseases. The role of the FOXM1-MMB complex in Wee1-like protein kinase (WEE1) inhibitor sensitivity in NSCLC keeps unclear. Methods: The reverse transcription quantitative polymerase chain reaction (RT-qPCR) was performed to measure the mRNA levels of FOXM1, LIN54, Replication Protein A (RPA), gammaH2AX (γH2AX) and Cyclin B (CCNB). The western blot was performed to examine the corresponding protein expressions. The Cell Counting Kit-8 (CCK-8) assay was performed to test cell survival. Result: It was demonstrated that after AZD-1775 treatment, the decrease in cell survival mediated by FOXM1 overexpression (P<0.001) could be reversed by LIN54 knockdown (P<0.01) and that cell survival in the control group did not differ obviously from that in the pcDNA3.1-FOXM1+siLIN54 group, indicating that the FOXM1-MMB complex was necessary for WEE1 inhibitor sensitivity. Moreover, the mRNA and protein expression levels of RPA and γH2AX were increased after AZD-1775 treatment and FOXM1 overexpression (P<0.01), suggesting that FOXM1 upregulation enhanced DNA replication stress and DNA damage. Finally, we found that the increases in the mRNA and protein expression levels of CCNB mediated by FOXM1 (P<0.01) could be rescued by silencing LIN54 (P<0.001) and that CCNB expression in the control group did not differ obviously from that in the pcDNA3.1-FOXM1+siLIN54 group. These findings revealed that the FOXM1-MMB complex activated G2/M checkpoints. In our work, it was discovered that FOXM1 overexpression increased DNA replication stress, which increased DNA replication and pressure on the WEE1 checkpoint. On the other hand, FOXM1 can enhance CCNB expression, increase the threshold content of the CCNB/CDK1 complex, facilitate mitosis, and promote WEE1 dephosphorylation. Under these two conditions, sensitivity to the WEE1 inhibitor AZD-1775 is increased, which leads to the accumulation of DNA damage and drives the activation of apoptosis. Conclusions: Overexpressed FOXM1 collaborates with MMB to increase WEE1 inhibitor sensitivity in NSCLC. This discovery might highlight the regulatory function of FOXM1/MMB in the treatment of NSCLC patients.
RESUMEN
INTRODUCTION: To investigate the diagnostic value of loop-mediated thermostatic amplification (LAMP) in detecting pathogenic bacteria from bronchoalveolar lavage fluid (BALF) of patients with pulmonary disorders combined with lower respiratory tract infections (LRTI). METHODOLOGY: This cross-sectional study included patients with pulmonary disorders combined with LRTI, including chronic obstructive pulmonary disease (COPD), bronchiectasis, or lung cancer, hospitalized in Meizhou People's Hospital between January 2020 and October 2021. BALF was collected using local bronchoalveolar lavage and electronic bronchoscopy. The presence of the pathogens was confirmed using the LAMP method and the bacterial culture method. RESULTS: In total, 249 patients were included (135 with COPD, 73 with bronchiectasis, and 41 with lung cancer). The proportions of Methicillin-resistant Staphylococcus aureus (4.8% vs 0.4%, p = 0.02) and Haemophilus influenzae (6.8% vs 0.4%, p < 0.001) detected by the LAMP method was higher, while the proportion of Pseudomonas aeruginosa was lower compared with that of the culture method (6.8% vs 12.4%, p = 0.034). The bacterial species with the highest agreement coefficient was Stenotrophomonas maltophilia (Kappa = 0.798, p < 0.001). Furthermore, 9 COPD patients exhibited mixed infections as determined by the LAMP method, whereas the culture method detected only 2 of these cases (1.48%) (p < 0.05). CONCLUSIONS: LAMP can detect more pathogenic bacteria, notably Haemophilus influenza, Methicillin-resistant Staphylococcus aureus, and atypical pathogens in patients with clinically common pulmonary disorders combined with LRTI. LAMP may provide etiological evidence to guide the clinical use of antibiotics in primary hospitals.
Asunto(s)
Bronquiectasia , Neoplasias Pulmonares , Staphylococcus aureus Resistente a Meticilina , Enfermedad Pulmonar Obstructiva Crónica , Infecciones del Sistema Respiratorio , Humanos , Staphylococcus aureus Resistente a Meticilina/genética , Estudios Transversales , Infecciones del Sistema Respiratorio/diagnóstico , Infecciones del Sistema Respiratorio/microbiología , Bacterias/genética , Enfermedad Pulmonar Obstructiva Crónica/diagnósticoRESUMEN
BACKGROUND: To determine the safety of Botox and its potential effect on alleviating radiation therapy (RT)-induced sialadenitis in head and neck cancer patients. METHODS: Twenty patients with stage III/IV head and neck cancer were randomized to receive Botox or saline injections into both submandibular glands (SMG). There were three visits: one before RT (V1); 1 week after RT (V2); and 6 weeks after RT (V3), each of which included saliva collection, a 24-h dietary recall, and a quality-of-life survey. RESULTS: No adverse events were observed. While the control group was much older, the Botox group more commonly underwent induction chemotherapy compared with controls. From V1 to V2, salivary flow decreased in both groups, but only in the control group from V1 to V3. CXCL-1 (GRO), a neutrophil chemoattractant, was lower in the Botox group compared with the control group at V3. CONCLUSION: Botox can be safely administered to the salivary glands prior to external beam radiation without observed complications or side-effects. After an initial reduction in salivary flow following RT, the Botox group showed lack of further flow reduction compared with controls. The inflammatory marker CXCL 1, which was reduced in the in Botox group at V3, may be a candidate for further studies of radiation-induced sialadenitis.